LOCUS       sequence-349375-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human APOE at the C-terminus 
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4377)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4377)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: APOE_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(533..1324,1325..1393))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1156..1175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1394..1498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1566..1584
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1622..1644)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1744..1763
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1957..1979
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1971..1988
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1972..1988
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2440..3327
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     primer_bind     complement(3749..3765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3749..3765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3762..3784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3773..3789
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3797..3803,3804..3821,3822..3827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3842..3863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3980..3997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4151..4377,1..362))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4231..4250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
       61 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
      601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
      661 caatgatacc gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag
      721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
      781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
      841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
      901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
      961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1501 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     1561 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     1621 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     1681 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     1741 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     1801 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     1861 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     1921 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     1981 cggccagtga attcccgcga gggcgccgag cgcggcctca gcgccatccg cgagcgcctg
     2041 gggcccctgg tggaacaggg ccgcgtgcgg gccgccactg tgggctccct ggccggccag
     2101 ccgctacagg agcgggccca ggcctggggc gagcggctgc gcgcgcggat ggaggagatg
     2161 ggcagccgga cccgcgaccg cctggacgag gtgaaggagc aggtggcgga ggtgcgcgcc
     2221 aagctggagg agcaggccca gcagatacgc ctgcaggccg aggccttcca ggcccgcctc
     2281 aagagctggt tcgagcccct ggtggaagac atgcagcgcc agtgggccgg gctggtggag
     2341 aaggtgcagg ctgccgtggg caccagcgcc gcccctgtgc ccagcgacaa tcacggtgga
     2401 ggaggttctg gaggcggtgg aagtggtggc ggaggtagcg ccgagatcgg caccggcttt
     2461 cccttcgatc ctcactacgt ggaagtgctg ggcgagagaa tgcactatgt ggacgtgggc
     2521 cccagagatg gaacccctgt gctgtttctg cacggcaacc ctaccagcag ctacgtgtgg
     2581 cggaacatca tccctcacgt ggcccctaca cacagatgta tcgcccctga cctgatcggc
     2641 atgggcaaga gcgataagcc cgacctgggc tacttcttcg acgaccacgt gcggttcatg
     2701 gacgccttta ttgaggccct cggcctggaa gaggtggtgc tggttattca cgattggggc
     2761 agcgccctgg gattccactg ggccaagaga aaccccgaga gagtgaaggg aatcgccttc
     2821 atggaattca tcaggcccat tcctacctgg gacgagtggc ctgagttcgc cagagagaca
     2881 ttccaggcct tcagaaccac cgacgtgggc agaaagctga tcatcgacca gaacgtgttc
     2941 atcgagggca ccctgcctat gggagtcgtc agaccactga ccgaggtgga aatggaccac
     3001 tacagagagc cctttctgaa ccccgtggac cgggaacctc tttggagatt ccccaacgag
     3061 ctgcccattg ccggcgagcc tgctaatatt gtggccctgg tggaagagta catggactgg
     3121 ctgcatcaga gccccgtgcc taagctgctg ttttggggaa cacccggcgt gctgattcct
     3181 cctgctgaag ctgccagact ggccaagagc ctgcctaatt gcaaggccgt ggatatcggc
     3241 cctggcctga atctgctgca agaggacaac cccgatctga tcggctccga gattgccaga
     3301 tggctgagca ccctggaaat cagcggataa acgccgaagc ctgcagccat gcgaccccac
     3361 gccaccccgt gcctcctgcc tccgcgcagc ctgcagcggg agaccctgtc cccgccccag
     3421 ccgtcctcct ggggtggacc ctagtttaat aaagattcac caagtttcac gcatctgctg
     3481 gcctccccct gtgatttcct ctaagcccca gcctcagttt ctctttctgc ccacatactg
     3541 gccacacaat tctcagcccc ctcctctcca tctgtgtctg tgtgtatctt tctctctgcc
     3601 cttttttttt tttttagacg gagtctggct ctgtcaccca ggctagagtg cagtggcacg
     3661 atcttggctc actgcaacct ctgcctcttg ggttcaagcg attctgctgc ctcagtagct
     3721 gggattacag aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3781 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3841 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3901 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3961 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4021 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4081 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4141 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4321 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgt
//
